RNA interference and receptors

@article{Goto2021,

title = {Verloren negatively regulates the expression of IMD pathway dependent antimicrobial peptides in Drosophila},

author = {Pragya Prakash and Arghyashree Roychowdhury-Sinha and Akira Goto},

url = {https://www.nature.com/articles/s41598-021-94973-0},

doi = {10.1038/s41598-021-94973-0},

year  = {2021},

date = {2021-07-30},

journal = {Scientific Reports},

volume = {11},

number = {15549},

abstract = {Drosophila immune deficiency (IMD) pathway is similar to the human tumor necrosis factor receptor (TNFR) signaling pathway and is preferentially activated by Gram-negative bacterial infection. Recent studies highlighted the importance of IMD pathway regulation as it is tightly controlled by numbers of negative regulators at multiple levels. Here, we report a new negative regulator of the IMD pathway, Verloren (Velo). Silencing of Velo led to constitutive expression of the IMD pathway dependent antimicrobial peptides (AMPs), and Escherichia coli stimulation further enhanced the AMP expression. Epistatic analysis indicated that Velo knock-down mediated AMP upregulation is dependent on the canonical members of the IMD pathway. The immune fluorescent study using overexpression constructs revealed that Velo resides both in the nucleus and cytoplasm, but the majority (~ 75%) is localized in the nucleus. We also observed from in vivo analysis that Velo knock-down flies exhibit significant upregulation of the AMP expression and reduced bacterial load. Survival experiments showed that Velo knock-down flies have a short lifespan and are susceptible to the infection of pathogenic Gram-negative bacteria, P. aeruginosa. Taken together, these data suggest that Velo is an additional new negative regulator of the IMD pathway, possibly acting in both the nucleus and cytoplasm.},

keywords = {bacteria, Biochemistry, DNA, Fungi, Gene Expression, gene regulation, Genetics, hoffmann, Immunochemistry, Immunology, infection, inflammation, Innate immune cells, innate immunity, M3i, microbiology, Molecular Biology, pathogens, RNA, RNAi, Signal Transduction, Transcription},

pubstate = {published},

tppubtype = {article}

}

@article{kobayashi_shigella_2013,

title = {The Shigella OspC3 effector inhibits caspase-4, antagonizes inflammatory cell death, and promotes epithelial infection},

author = {Taira Kobayashi and Michinaga Ogawa and Takahito Sanada and Hitomi Mimuro and Minsoo Kim and Hiroshi Ashida and Reiko Akakura and Mitsutaka Yoshida and Magdalena Kawalec and Jean-Marc Reichhart and Tsunehiro Mizushima and Chihiro Sasakawa},

doi = {10.1016/j.chom.2013.04.012},

issn = {1934-6069},

year  = {2013},

date = {2013-05-01},

journal = {Cell Host Microbe},

volume = {13},

number = {5},

pages = {570--583},

abstract = {Caspase-mediated inflammatory cell death acts as an intrinsic defense mechanism against infection. Bacterial pathogens deploy countermeasures against inflammatory cell death, but the mechanisms by which they do this remain largely unclear. In a screen for Shigella flexneri effectors that regulate cell death during infection, we discovered that Shigella infection induced acute inflammatory, caspase-4-dependent epithelial cell death, which is counteracted by the bacterial OspC3 effector. OspC3 interacts with the caspase-4-p19 subunit and inhibits its activation by preventing caspase-4-p19 and caspase-4-p10 heterodimerization by depositing the conserved OspC3 X1-Y-X₂-D-X₃ motif at the putative catalytic pocket of caspase-4. Infection of guinea pigs with a Shigella ospC3-deficient mutant resulted in enhanced inflammatory cell death and associated symptoms, correlating with decreased bacterial burdens. Salmonella Typhimurium and enteropathogenic Escherichia coli infection also induced caspase-4-dependent epithelial death. These findings highlight the importance of caspase-4-dependent innate immune responses and demonstrate that Shigella delivers a caspase-4-specific inhibitor to delay epithelial cell death and promote infection.},

keywords = {Animal, Animals, Bacillary, Bacterial, Bacterial Proteins, Caspases, Cell Death, Cell Line, Disease Models, DNA, Dysentery, Enzyme Inhibitors, Epithelial Cells, Escherichia coli, Gene Knockout Techniques, Guinea Pigs, Host-Pathogen Interactions, Humans, Initiator, M3i, Protein Binding, Protein Interaction Mapping, reichhart, Salmonella typhimurium, Sequence Analysis, Shigella flexneri, Virulence Factors},

pubstate = {published},

tppubtype = {article}

}

@article{deleury_specific_2012,

title = {Specific versus non-specific immune responses in an invertebrate species evidenced by a comparative de novo sequencing study},

author = {Emeline Deleury and Géraldine Dubreuil and Namasivayam Elangovan and Eric Wajnberg and Jean-Marc Reichhart and Benjamin Gourbal and David Duval and Olga Lucia Baron and Jérôme Gouzy and Christine Coustau},

doi = {10.1371/journal.pone.0032512},

issn = {1932-6203},

year  = {2012},

date = {2012-01-01},

journal = {PLoS ONE},

volume = {7},

number = {3},

pages = {e32512},

abstract = {Our present understanding of the functioning and evolutionary history of invertebrate innate immunity derives mostly from studies on a few model species belonging to ecdysozoa. In particular, the characterization of signaling pathways dedicated to specific responses towards fungi and Gram-positive or Gram-negative bacteria in Drosophila melanogaster challenged our original view of a non-specific immunity in invertebrates. However, much remains to be elucidated from lophotrochozoan species. To investigate the global specificity of the immune response in the fresh-water snail Biomphalaria glabrata, we used massive Illumina sequencing of 5'-end cDNAs to compare expression profiles after challenge by Gram-positive or Gram-negative bacteria or after a yeast challenge. 5'-end cDNA sequencing of the libraries yielded over 12 millions high quality reads. To link these short reads to expressed genes, we prepared a reference transcriptomic database through automatic assembly and annotation of the 758,510 redundant sequences (ESTs, mRNAs) of B. glabrata available in public databases. Computational analysis of Illumina reads followed by multivariate analyses allowed identification of 1685 candidate transcripts differentially expressed after an immune challenge, with a two fold ratio between transcripts showing a challenge-specific expression versus a lower or non-specific differential expression. Differential expression has been validated using quantitative PCR for a subset of randomly selected candidates. Predicted functions of annotated candidates (approx. 700 unisequences) belonged to a large extend to similar functional categories or protein types. This work significantly expands upon previous gene discovery and expression studies on B. glabrata and suggests that responses to various pathogens may involve similar immune processes or signaling pathways but different genes belonging to multigenic families. These results raise the question of the importance of gene duplication and acquisition of paralog functional diversity in the evolution of specific invertebrate immune responses.},

keywords = {Animals, Biomphalaria, Calmodulin, Cluster Analysis, Complementary, DNA, Expressed Sequence Tags, Ferritins, Gene Expression Profiling, Gene Expression Regulation, High-Throughput Nucleotide Sequencing, Immunity, Innate, M3i, messenger, Pattern Recognition, Phylogeny, Receptors, reichhart, RNA, Signal Transduction, Zinc Fingers},

pubstate = {published},

tppubtype = {article}

}

@article{niehus_fly_2012b,

title = {Fly culture collapse disorder: detection, prophylaxis and eradication of the microsporidian parasite Tubulinosema ratisbonensis infecting Drosophila melanogaster},

author = {Sebastian Niehus and Philippe Giammarinaro and Samuel Liégeois and Jessica Quintin and Dominique Ferrandon},

doi = {10.4161/fly.20896},

issn = {1933-6942},

year  = {2012},

date = {2012-01-01},

journal = {Fly (Austin)},

volume = {6},

number = {3},

pages = {193--204},

abstract = {Drosophila melanogaster is a robust model to investigate many biological problems. It is however prone to some infections, which may endanger fly stocks if left unchecked for. One such infection is caused by an obligate fungal intracellular parasite, Tubulinosema ratisbonensis, which can be found in laboratory stocks. Here, we identify and briefly characterize a T. ratisbonensis strain that was infesting our Drosophila cultures and that required intensive measures to contain and eradicate the infection. We describe the phenotypes of infested stocks. We also report PCR-based techniques that allow the detection of infested stocks with a high sensitivity. We have developed a high-throughput qPCR assay that allows the efficient parallel screening of a large number of potentially-infested stocks. We also have investigated several prophylactic measures to prevent the further contamination of stocks, namely UV-exposure, ethanol treatment, bleaching, and desiccation. Bleaching was found to kill all spores. Other treatments were less effective but were found to be sufficient to prevent further contamination of noninfested stocks. Two treatments were efficacious in curing infested stocks (1) bleaching of eggs and subsequent raising of the larvae in clean vials; (2) fumagillin treatment. These cures only work on stocks that have not become too weak to withstand the procedures.},

keywords = {Animals, Apansporoblastina, Apansporoblastina/*genetics/physiology, Base Sequence, cure, Disinfection, Disinfection/methods, DNA, DNA Primers, Drosophila melanogaster/*microbiology, ferrandon, fumagillin, Fungal, Fungal/chemistry, M3i, microsporidia, obligate intracellular parasitism, PCR detection, Phylogeny, Polymerase Chain Reaction, Polymerase Chain Reaction/methods, prophylaxis, Ribosomal, Ribosomal/chemistry, Sequence Alignment, Tubulinosema ratisbonensis},

pubstate = {published},

tppubtype = {article}

}

@article{schickel_carabin_2012,

title = {Carabin deficiency in B cells increases BCR-TLR9 costimulation-induced autoimmunity},

author = {Jean-Nicolas Schickel and Jean-Louis Pasquali and Anne Soley and Anne-Marie Knapp and Marion Decossas and Aurélie Kern and Jean-Daniel Fauny and Luc Marcellin and Anne-Sophie Korganow and Thierry Martin and Pauline Soulas-Sprauel},

doi = {10.1002/emmm.201201595},

issn = {1757-4684},

year  = {2012},

date = {2012-01-01},

journal = {EMBO molecular medicine},

volume = {4},

number = {12},

pages = {1261--1275},

abstract = {The mechanisms behind flares of human autoimmune diseases in general, and of systemic lupus in particular, are poorly understood. The present scenario proposes that predisposing gene defects favour clinical flares under the influence of external stimuli. Here, we show that Carabin is low in B cells of (NZB × NZW) F1 mice (murine SLE model) long before the disease onset, and is low in B cells of lupus patients during the inactive phases of the disease. Using knock-out and B-cell-conditional knock-out murine models, we identify Carabin as a new negative regulator of B-cell function, whose deficiency in B cells speeds up early B-cell responses and makes the mice more susceptible to anti-dsDNA production and renal lupus flare after stimulation with a Toll-like Receptor 9 agonist, CpG-DNA. Finally, in vitro analysis of NFκB activation and Erk phosphorylation in TLR9- and B-cell receptor (BCR)-stimulated Carabin-deficient B cells strongly suggests how the internal defect synergizes with the external stimulus and proposes Carabin as a natural inhibitor of the potentially dangerous crosstalk between BCR and TLR9 pathways in self-reactive B cells.},

keywords = {Adaptor Proteins, Animals, Antigen, Autoimmunity, B-Cell, B-Lymphocytes, Carrier Proteins, Cohort Studies, DNA, Humans, I2CT, Imagerie, Inbred NZB, Inbred Strains, Mice, Phosphorylation, Prospective Studies, Receptors, Signal Transducing, Toll-Like Receptor 9, Transfection},

pubstate = {published},

tppubtype = {article}

}

@article{montellano_fullerene_2011,

title = {Fullerene C₆₀ as a multifunctional system for drug and gene delivery},

author = {Alejandro Montellano and Tatiana Da Ros and Alberto Bianco and Maurizio Prato},

doi = {10.1039/c1nr10783f},

issn = {2040-3372},

year  = {2011},

date = {2011-10-01},

journal = {Nanoscale},

volume = {3},

number = {10},

pages = {4035--4041},

abstract = {The fullerene family, and especially C(60), has delighted the scientific community during the last 25 years with perspective applications in a wide variety of fields, including the biological and the biomedical domains. Several biomedical uses have been explored using water-soluble C(60)-derivatives. However, the employment of fullerenes for drug delivery is still at an early stage of development. The design and synthesis of multifunctionalized and multimodal C(60) systems able to cross the cell membranes and efficiently deliver active molecules is an attracting challenge that involves multidisciplinary strategies. Promising results have emerged in the last years, bringing fullerenes again to the front of interest. Herein, the state of the art of this emerging field is presented and illustrated with some of the most representative examples.},

keywords = {DNA, Drug Carriers, Fullerenes, Gene Transfer Techniques, I2CT, Immunoconjugates, Plasmids, Team-Bianco},

pubstate = {published},

tppubtype = {article}

}

@article{bou_aoun_analysis_2011,

title = {Analysis of thioester-containing proteins during the innate immune response of Drosophila melanogaster},

author = {Richard Bou Aoun and Charles Hetru and Laurent Troxler and Daniel Doucet and Dominique Ferrandon and Nicolas Matt},

doi = {10.1159/000321554},

issn = {1662-8128},

year  = {2011},

date = {2011-01-01},

journal = {J Innate Immun},

volume = {3},

number = {1},

pages = {52--64},

abstract = {Thioester-containing proteins (TEPs) are conserved proteins among insects that are thought to be involved in innate immunity. In Drosophila, the Tep family is composed of 6 genes named Tep1-Tep6. In this study, we investigated the phylogeny, expression pattern and roles of these genes in the host defense of Drosophila. Protostomian Tep genes are clustered in 3 distinct branches, 1 of which is specific to mosquitoes. Most D. melanogaster Tep genes are expressed in hemocytes, can be induced in the fat body, and are expressed in specific regions of the hypodermis. This expression pattern is consistent with a role in innate immunity. However, we find that TEP1, TEP2, and TEP4 are not strictly required in the body cavity to fight several bacterial and fungal infections. One possibility is that Drosophila TEPs act redundantly or that their absence can be compensated by other components of the immune response. TEPs may thus provide a subtle selective advantage during evolution. Alternatively, they may be required in host defense against specific as yet unidentified natural pathogens of Drosophila.},

keywords = {Animals, bioinformatic, DNA, Evolution, ferrandon, Gene Expression Regulation, Hemocytes, Immunity, In Situ Hybridization, Innate, M3i, matt, Molecular, Mutation, Phylogeny, Sequence Analysis},

pubstate = {published},

tppubtype = {article}

}

@article{,

title = {Kinetics of deoxy-CTP incorporation opposite a dG-C8-N-2-aminofluorene adduct by a high-fidelity DNA polymerase},

author = { D. Y. Burnouf and J. E. Wagner},

year  = {2009},

date = {2009-01-01},

journal = {J Mol Biol},

volume = {386},

number = {4},

pages = {951-61},

abstract = {The model carcinogen N-2-acetylaminofluorene covalently binds to the C8 position of guanine to form two adducts, the N-(2'-deoxyguanosine-8-yl)-aminofluorene (G-AF) and the N-2-(2'-deoxyguanosine-8-yl)-acetylaminofluorene (G-AAF). Although they are chemically closely related, their biological effects are strongly different and they are processed by different damage tolerance pathways. G-AF is bypassed by replicative and high-fidelity polymerases, while specialized polymerases ensure synthesis past of G-AAF. We used the DNA polymerase I fragment of a Bacillus stearothermophilus strain as a model for a high-fidelity polymerase to study the kinetics of incorporation of deoxy-CTP (dCTP) opposite a single G-AF. Pre-steady-state kinetic experiments revealed a drastic reduction in dCTP incorporation performed by the G-AF-modified ternary complex. Two populations of these ternary complexes were identified: (i) a minor productive fraction (20%) that readily incorporates dCTP opposite the G-AF adduct with a rate similar to that measured for the adduct-free ternary complexes and (ii) a major fraction of unproductive complexes (80%) that slowly evolve into productive ones. In the light of structural data, we suggest that this slow rate reflects the translocation of the modified base within the active site, from the pre-insertion site into the insertion site. By making this translocation rate limiting, the G-AF lesion reveals a novel kinetic step occurring after dNTP binding and before chemistry.},

note = {1089-8638 (Electronic) 

Journal Article 

Research Support, Non-U.S. Gov't},

keywords = {Adducts, Bacillus, Catalytic, Cytidine, Deoxyguanosine/*metabolism, DNA, DNA-Directed, Domain, DUMAS, Elements, Fluorenes/*metabolism, Guanine, Kinetics, Oligonucleotides/metabolism, Phosphorothioate, Polymerase/*metabolism, Specificity, stearothermophilus/enzymology, Substrate, Titrimetry, Triphosphate/*metabolism},

pubstate = {published},

tppubtype = {article}

}

@article{garrett_identification_2009,

title = {Identification and analysis of serpin-family genes by homology and synteny across the 12 sequenced Drosophilid genomes},

author = {Matthew Garrett and Ane Fullaondo and Laurent Troxler and Gos Micklem and David Gubb},

doi = {10.1186/1471-2164-10-489},

issn = {1471-2164},

year  = {2009},

date = {2009-01-01},

journal = {BMC Genomics},

volume = {10},

pages = {489},

abstract = {BACKGROUND: The Drosophila melanogaster genome contains 29 serpin genes, 12 as single transcripts and 17 within 6 gene clusters. Many of these serpins have a conserved "hinge" motif characteristic of active proteinase inhibitors. However, a substantial proportion (42%) lacks this motif and represents non-inhibitory serpin-fold proteins of unknown function. Currently, it is not known whether orthologous, inhibitory serpin genes retain the same target proteinase specificity within the Drosophilid lineage, nor whether they give rise to non-inhibitory serpin-fold proteins or other, more diverged, proteins. RESULTS: We collated 188 orthologues to the D. melanogaster serpins from the other 11 Drosophilid genomes and used synteny to find further family members, raising the total to 226, or 71% of the number of orthologues expected assuming complete conservation across all 12 Drosophilid species. In general the sequence constraints on the serpin-fold itself are loose. The critical Reactive Centre Loop (RCL) sequence, including the target proteinase cleavage site, is strongly conserved in inhibitory serpins, although there are 3 exceptional sets of orthologues in which the evolutionary constraints are looser. Conversely, the RCL of non-inhibitory serpin orthologues is less conserved, with 3 exceptions that presumably bind to conserved partner molecules. We derive a consensus hinge motif, for Drosophilid inhibitory serpins, which differs somewhat from that of the vertebrate consensus. Three gene clusters appear to have originated in the melanogaster subgroup, Spn28D, Spn77B and Spn88E, each containing one inhibitory serpin orthologue that is present in all Drosophilids. In addition, the Spn100A transcript appears to represent a novel serpin-derived fold. CONCLUSION: In general, inhibitory serpins rarely change their range of proteinase targets, except by a duplication/divergence mechanism. Non-inhibitory serpins appear to derive from inhibitory serpins, but not the reverse. The conservation of different family members varied widely across the 12 sequenced Drosophilid genomes. An approach considering synteny as well as homology was important to find the largest set of orthologues.},

keywords = {Animals, bioinformatic, Comparative Genomic Hybridization, Conserved Sequence, DNA, Drosophilidae, Evolution, Genome, Insect, Molecular, Multigene Family, Sequence Alignment, Sequence Analysis, Serpins, Synteny},

pubstate = {published},

tppubtype = {article}

}

@article{muller_spliceosomal_2008,

title = {Spliceosomal peptide P140 for immunotherapy of systemic lupus erythematosus: results of an early phase II clinical trial},

author = {Sylviane Muller and Fanny Monneaux and Nicolas Schall and Rasho K Rashkov and Boycho A Oparanov and Philippe Wiesel and Jean-Marie Geiger and Robert Zimmer},

doi = {10.1002/art.24027},

issn = {0004-3591},

year  = {2008},

date = {2008-01-01},

journal = {Arthritis and Rheumatism},

volume = {58},

number = {12},

pages = {3873--3883},

abstract = {OBJECTIVE: To assess the safety, tolerability, and efficacy of spliceosomal peptide P140 (IPP-201101; sequence 131-151 of the U1-70K protein phosphorylated at Ser140), which is recognized by lupus CD4+ T cells, in the treatment of patients with systemic lupus erythematosus (SLE). 

METHODS: An open-label, dose-escalation phase II study was conducted in two centers in Bulgaria. Twenty patients (2 male and 18 female) with moderately active SLE received 3 subcutaneous (SC) administrations of a clinical batch of P140 peptide at 2-week intervals. Clinical evaluation was performed using approved scales. A panel of autoantibodies, including antinuclear antibodies, antibodies to extractable nuclear antigens (U1 RNP, SmD1, Ro/SSA, La/SSB), and antibodies to double-stranded DNA (anti-dsDNA), chromatin, cardiolipin, and peptides of the U1-70K protein, was tested by enzyme-linked immunosorbent assay (ELISA). The plasma levels of C-reactive protein, total Ig, IgG, IgG subclasses, IgM, IgA, and IgE, and of the cytokines interleukin-2 and tumor necrosis factor alpha were measured by ELISA and nephelometry. 

RESULTS: IgG anti-dsDNA antibody levels decreased by at least 20% in 7 of 10 patients who received 3 x 200 microg IPP-201101 (group 1), but only in 1 patient in the group receiving 3 x 1,000 microg IPP-201101 (group 2). Physician's global assessment of disease activity scores and scores on the SLE Disease Activity Index were significantly decreased in group 1. The changes occurred progressively in the population of responders, increased in magnitude during the treatment period, and were sustained. No clinical or biologic adverse effects were observed in the individuals, except for some local irritation at the highest concentration. 

CONCLUSION: IPP-201101 was found to be safe and well tolerated by subjects. Three SC doses of IPP-201101 at 200 microg significantly improved the clinical and biologic status of lupus patients.},

keywords = {Adolescent, Adult, Aged, Antibodies, Antinuclear, C-Reactive Protein, DNA, Female, Humans, I2CT, Immunotherapy, Lupus Erythematosus, Male, Middle Aged, Monneaux, Peptide Fragments, Peptides, Severity of Illness Index, Spliceosomes, Systemic, Team-Dumortier, Treatment Outcome, Young Adult},

pubstate = {published},

tppubtype = {article}

}

@article{,

title = {Identification of new noncoding RNAs in Listeria monocytogenes and prediction of mRNA targets},

author = { P. Mandin and F. Repoila and M. Vergassola and T. Geissmann and P. Cossart},

year  = {2007},

date = {2007-01-01},

journal = {Nucleic Acids Res},

volume = {35},

number = {3},

pages = {962-74},

abstract = {To identify noncoding RNAs (ncRNAs) in the pathogenic bacterium Listeria monocytogenes, we analyzed the intergenic regions (IGRs) of strain EGD-e by in silico-based approaches. Among the twelve ncRNAs found, nine are novel and specific to the Listeria genus, and two of these ncRNAs are expressed in a growth-dependent manner. Three of the ncRNAs are transcribed in opposite direction to overlapping open reading frames (ORFs), suggesting that they act as antisense on the corresponding mRNAs. The other ncRNA genes appear as single transcription units. One of them displays five repeats of 29 nucleotides. Five of these new ncRNAs are absent from the non-pathogenic species L. innocua, raising the possibility that they might be involved in virulence. To predict mRNA targets of the ncRNAs, we developed a computational method based on thermodynamic pairing energies and known ncRNA-mRNA hybrids. Three ncRNAs, including one of the putative antisense ncRNAs, were predicted to have more than one mRNA targets. Several of them were shown to bind efficiently to the ncRNAs suggesting that our in silico approach could be used as a general tool to search for mRNA targets of ncRNAs.},

note = {1362-4962 (Electronic) 

Journal Article 

Research Support, Non-U.S. Gov't},

keywords = {5', Assay, Bacterial, Base, Biology, Computational, Data, DNA, Electrophoretic, Flanking, Genes, Genomics, Intergenic/chemistry, Listeria, Messenger/chemistry/*metabolism, Mobility, Molecular, monocytogenes/*genetics/metabolism, Region, RNA, ROMBY, Sequence, Shift, Untranslated/analysis/*genetics/metabolism},

pubstate = {published},

tppubtype = {article}

}

@article{monneaux_spliceosome_2007,

title = {[The spliceosome and its interest for lupus therapy]},

author = {F Monneaux and S Muller},

doi = {10.1016/j.revmed.2007.05.003},

issn = {0248-8663},

year  = {2007},

date = {2007-01-01},

journal = {La Revue De Medecine Interne},

volume = {28},

number = {10},

pages = {725--728},

abstract = {INTRODUCTION: The spliceosome, which is a particle containing a molecule of U-RNA and proteins that are specific to each U ribonuclear particle (U-snRNP) or common to every U-snRNPs, is one of the numerous nuclear targets recognized by the antibodies (Abs) and CD4+ T cells from patients with systemic lupus erythematosus and lupus mice. 

EXEGESIS: We recently characterized a peptide from the spliceosomal protein U1-70K (sequence 131-151), which is recognized by the Abs and CD4+ T cells from lupus mice and patients. This peptide contains a conserved RNP1 motif, which is also present in other spliceosomal proteins targeted by the Abs from individuals with lupus. We further showed that peptide 131-151 containing a phosphoserine at position 140 (peptide P140) possessed tolerogenic properties in lupus mice and was recognized by the Abs and CD4+ T cells from lupus patients. 

CONCLUSION: Thanks to its RNP1 motif, the peptide P140 might play an important role in the initiation and perpetuation steps of the humoral and cellular immune response diversification in lupus individuals. Therapeutic and particularly immunomodulating properties of P140 peptide are being evaluated in humans (a phase III clinical trial will be undertaken in the next weeks).},

keywords = {Amino Acid Motifs, Animals, Antibodies, CD4-Positive T-Lymphocytes, Conserved Sequence, DNA, Epitopes, Haplotypes, Humans, I2CT, Immune Tolerance, Inbred MRL lpr, Inbred NZB, Lupus Erythematosus, Mice, Monneaux, Phosphoserine, Protein, Recombinant, Ribonucleoprotein, Sequence Analysis, Serine, Spliceosomes, Systemic, Team-Dumortier, U1 Small Nuclear},

pubstate = {published},

tppubtype = {article}

}

@article{klumpp_multifunctionalised_2007,

title = {Multifunctionalised cationic fullerene adducts for gene transfer: design, synthesis and DNA complexation},

author = {Cédric Klumpp and Lara Lacerda and Olivier Chaloin and Tatiana Da Ros and Kostas Kostarelos and Maurizio Prato and Alberto Bianco},

doi = {10.1039/b708435h},

issn = {1359-7345},

year  = {2007},

date = {2007-01-01},

journal = {Chemical Communications (Cambridge, England)},

number = {36},

pages = {3762--3764},

abstract = {Cationic poly-N,N-dimethylfulleropyrrolidinium derivatives have been designed and synthesised to complex plasmid DNA for gene delivery.},

keywords = {DNA, Electrophoresis, Fullerenes, Gene Transfer Techniques, I2CT, Molecular Structure, Plasmids, Team-Bianco},

pubstate = {published},

tppubtype = {article}

}

@article{klumpp_functionalized_2006,

title = {Functionalized carbon nanotubes as emerging nanovectors for the delivery of therapeutics},

author = {Cédric Klumpp and Kostas Kostarelos and Maurizio Prato and Alberto Bianco},

doi = {10.1016/j.bbamem.2005.10.008},

issn = {0006-3002},

year  = {2006},

date = {2006-03-01},

journal = {Biochimica Et Biophysica Acta},

volume = {1758},

number = {3},

pages = {404--412},

abstract = {Functionalized carbon nanotubes (f-CNT) are emerging as a new family of nanovectors for the delivery of different types of therapeutic molecules. The application of CNT in the field of carrier-mediated delivery has become possible after the recent discovery of their capacity to penetrate into the cells. CNT can be loaded with active molecules by forming stable covalent bonds or supramolecular assemblies based on noncovalent interactions. Once the cargos are carried into various cells, tissues and organs they are able to express their biological function. In this review, we will describe the potential of f-CNT to deliver different types of therapeutic molecules.},

keywords = {carbon, DNA, Drug Carriers, Drug Delivery Systems, Humans, I2CT, Nanotubes, RNA, Team-Bianco},

pubstate = {published},

tppubtype = {article}

}

@article{lacerda_luminescence_2006,

title = {Luminescence of Functionalized Carbon Nanotubes as a Tool to Monitor Bundle Formation and Dissociation in Water: The Effect of Plasmid-DNA Complexation},

author = {L Lacerda and G Pastorin and W Wu and M Prato and A Bianco and K Kostarelos},

url = {https://onlinelibrary.wiley.com/doi/abs/10.1002/adfm.200500569},

doi = {10.1002/adfm.200500569},

issn = {1616-3028},

year  = {2006},

date = {2006-01-01},

urldate = {2020-03-31},

journal = {Advanced Functional Materials},

volume = {16},

number = {14},

pages = {1839--1846},

abstract = {Functionalized carbon nanotubes (f-CNTs) are explored as novel nanomaterials for biomedical applications. UV-vis luminescence of aqueous dispersions of CNT–NH3+ and CNT–NH–Ac (NH–Ac: acetamido) is observed using standard laboratory spectrophotometric instrumentation, and the measured fluorescence intensity is correlated with the aggregation state of the f-CNTs: a high intensity indicates improved f-CNT individualization and dispersion, while a decrease in fluorescence intensity indicates a higher degree of nanotube aggregation and bundling as a result of varying the sodium dodecyl sulfate (SDS) concentrations and pH in the aqueous phase. Moreover, utilization of this relationship between fluorescence intensity and the state of f-CNT aggregation is carried out to elucidate the interactions between f-CNTs and gene-encoding plasmid DNA (pDNA). pDNA is shown to interact with CNT–NH3+ primarily through electrostatic interactions that lead concomitantly to a higher degree of f-CNT bundling. The CNT–NH3+/pDNA interactions are successfully competed by SDS/f-CNT surface interactions, resulting in the displacement of pDNA. These studies provide exemplification of the use of fluorescence spectrophotometry to accurately describe the aggregation state of water-soluble f-CNTs. Characterization of the complexes between pDNA and f-CNTs elucidates the opportunities and limitations of such supramolecular systems as potential vectors for gene transfer.},

keywords = {Carbon nanotubes, DNA, I2CT, Luminescence, multiwalled, single-walled, Team-Bianco},

pubstate = {published},

tppubtype = {article}

}

@article{bianco_carbon_2005,

title = {Carbon nanotubes: on the road to deliver},

author = {Alberto Bianco and Johan Hoebeke and Kostas Kostarelos and Maurizio Prato and Charalambos D Partidos},

doi = {10.2174/1567201054367959},

issn = {1567-2018},

year  = {2005},

date = {2005-07-01},

journal = {Current Drug Delivery},

volume = {2},

number = {3},

pages = {253--259},

abstract = {Over the last few years, considerable advances have been made in the field of nanotechnology. The advent of carbon nanotube functionalization has paved the way for their potential application as a delivery system of diverse molecules such as peptides, proteins, plasmid DNA, and synthetic oligodeoxynucleotides. This opens new therapeutic and preventive opportunities to combat diseases. The scope of this review is to summarize our recent work in this rapidly growing field.},

keywords = {carbon, CpG Islands, DNA, Drug Carriers, I2CT, nanotechnology, Team-Bianco},

pubstate = {published},

tppubtype = {article}

}

@article{singh_binding_2005,

title = {Binding and condensation of plasmid DNA onto functionalized carbon nanotubes: toward the construction of nanotube-based gene delivery vectors},

author = {Ravi Singh and Davide Pantarotto and David McCarthy and Olivier Chaloin and Johan Hoebeke and Charalambos D Partidos and Jean-Paul Briand and Maurizio Prato and Alberto Bianco and Kostas Kostarelos},

doi = {10.1021/ja0441561},

issn = {0002-7863},

year  = {2005},

date = {2005-03-01},

journal = {Journal of the American Chemical Society},

volume = {127},

number = {12},

pages = {4388--4396},

abstract = {Carbon nanotubes (CNTs) constitute a class of nanomaterials that possess characteristics suitable for a variety of possible applications. Their compatibility with aqueous environments has been made possible by the chemical functionalization of their surface, allowing for exploration of their interactions with biological components including mammalian cells. Functionalized CNTs (f-CNTs) are being intensively explored in advanced biotechnological applications ranging from molecular biosensors to cellular growth substrates. We have been exploring the potential of f-CNTs as delivery vehicles of biologically active molecules in view of possible biomedical applications, including vaccination and gene delivery. Recently we reported the capability of ammonium-functionalized single-walled CNTs to penetrate human and murine cells and facilitate the delivery of plasmid DNA leading to expression of marker genes. To optimize f-CNTs as gene delivery vehicles, it is essential to characterize their interactions with DNA. In the present report, we study the interactions of three types of f-CNTs, ammonium-functionalized single-walled and multiwalled carbon nanotubes (SWNT-NH3+; MWNT-NH3+), and lysine-functionalized single-walled carbon nanotubes (SWNT-Lys-NH3+), with plasmid DNA. Nanotube-DNA complexes were analyzed by scanning electron microscopy, surface plasmon resonance, PicoGreen dye exclusion, and agarose gel shift assay. The results indicate that all three types of cationic carbon nanotubes are able to condense DNA to varying degrees, indicating that both nanotube surface area and charge density are critical parameters that determine the interaction and electrostatic complex formation between f-CNTs with DNA. All three different f-CNT types in this study exhibited upregulation of marker gene expression over naked DNA using a mammalian (human) cell line. Differences in the levels of gene expression were correlated with the structural and biophysical data obtained for the f-CNT:DNA complexes to suggest that large surface area leading to very efficient DNA condensation is not necessary for effective gene transfer. However, it will require further investigation to determine whether the degree of binding and tight association between DNA and nanotubes is a desirable trait to increase gene expression efficiency in vitro or in vivo. This study constitutes the first thorough investigation into the physicochemical interactions between cationic functionalized carbon nanotubes and DNA toward construction of carbon nanotube-based gene transfer vector systems.},

keywords = {carbon, Cations, DNA, Electron, Gene Transfer Techniques, Genetic Vectors, I2CT, Lysine, Microscopy, Nanotubes, Plasmids, Quaternary Ammonium Compounds, scanning, Surface Plasmon Resonance, Team-Bianco},

pubstate = {published},

tppubtype = {article}

}